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Previous issue date: 2009 / Fundação Oswaldo Cruz.Instituto Oswaldo Cruz. Rio de janeiro, RJ, Brasil / No centenario da descoberta da doenca de Chagas (DC), esta parasitose negligenciada causada pelo Trypanosoma cruzi e importante causa de mortalidade e morbidade na America Latina, sem vacinas ou agentes quimioterapicos satisfatorios. Esta tese objetivou analisar atividade, seletividade e mecanismos de acao sobre o T.cruzi de 04 classes de compostos (fluorquinolonas - FQ, tiossemicarbazonas - TS, diamidinas aromaticas - DA e arilimidamidas – AIA – uma nova classe a partir de DA) in vitro e in vivo. FQ como NOR e SPAR sao antibioticos com ampla acao bactericida, atuando sobre o DNA via toposoimerases. Embora SPAR e NOR nao tenham sido ativas, seus complexos metalicos de cobre-fenantrolina foram efetivos sobre tripomastigotas e formas intracelulares de T.cruzi, (IC50 1,62 a 4,65µM). TS sao agentes anti-tumorais e microbicidas que atuam sobre ribonucleotideo redutases e cisteina proteases. Todas TS apresentaram baixa toxicidade (>200µM) sobre celulas de mamiferos. Embora N4-metil-4-nitrobenzaldeido-tiossemicarbazona, N4-metill-4-nitroacetofenona-tiossemicarbazona e seus complexos metalicos de manganes (Mn) não tenham sido ativos, o complexo de Mn da N4-metil-4-nitrobenzofenona-tiossemicarbazona revelou-se moderadamente ativo sobre formas sanguineas (IC50 19µM). A terceira classe foi DA, ligantes de DNA com excelente atividade sobre diversos patogenos. Embora todas tenham baixa toxicidade sobre cardiomiocitos, somente tres (DB1645, DB1582 e DB1651) foram ativas contra formas sanguineas e intracelulares (IC50 entre 0,15 a 13,3µM), com atividade relacionada a curvatura das moleculas (sendo as moleculas lineares as nao efetivas). Todas DA localizam-se no nucleo e kDNA dos parasitos, com maior acumulo na ulti
Analise in vitro da AIA DB766 sobre T.cruzi revelou: (i) excelente atividade sobre formas sanguineas (60nM) e intracelulares (25nM), mantendo otima acao mesmo quando o parasito e incubado com 96% de sangue de camundongo, sugerindo assim, um potencial uso profilatico, (ii) ação sobre diferentes cepas (peridomiciliares/silvestres), incluindo as naturalmente resistentes ao benznidazole (BZ), com superior eficacia que BZ, (iii) localizacao em compartimentos ricos em DNA, induzindo danos ultra-estruturais na mitocondria. Com base no excelente indice de seletividade (>533 e 714), ensaios foram conduzidos durante a infeccao aguda experimental por T.cruzi (cepas Y e Colombiana), em doses diarias ou alternadas =100mg/kg/dia da DB766 sozinha ou associada ao BZ, administradas via ip ou p.o. por ate 20 dias, com primeira dose no inicio da parasitemia. DB766 (25 e 50mg/kg/dia ip e 100mg/kg/dia p.o.) reduziu (>90%) a carga parasitaria (sangue e tecido cardiaco) e protegeu 90-100% contra mortalidade, com semelhante eficacia ao BZ. AIA reverteu inflamacao e alteracoes eletricas cardiacas e protegeu contra lesoes hepaticas e musculares induzidas pela infeccao. Doses sub-otimas de BZ associado com a DB289 (DA - pro-droga da DB75 – via p.o.) ou com a DB766 (ip) resultaram em =99% reducao de parasitemia e 100% de protecao sobre mortalidade. BZ (p.o)+DB766 (ip) resultou em cura parasitologica (2 em 15 animais) avaliada por hemocultivo e PCR. AIA foi a mais ativa e seletiva sobre T. cruzi, estimulando a continuidade de ensaios pre-clinicos com representantes desta classe visando identificacao de novos farmacos para a DC / In the centenary of the discovery of Chagas' disease (AD), this parasitosis caused by Trypanosoma cruzi neglected and important cause of morbidity and mortality in Latin America, no vaccines or chemotherapeutic agents satisfactory. This thesis aims to analyze activity, selectivity and mechanisms of action of T. cruzi on the 04 classes of compounds (fluoroquinolones - CF thiosemicarbazones - TS, aromatic diamidines - DA and arilimidamidas - AIA - a new class from DA) in vitro and in vivo. CF as NOR and SPAR are antibiotics with broad antibacterial action, acting on the DNA via toposoimerases. Although SPAR and NOR have not been active, their metallic complexes of copper-phenanthroline were effective on trypomastigotes and intracellular forms of T. cruzi (IC50 1.62 to 4.65 mM). TS are anti-tumor agents and microbicides that act on ribonucleotide reductase and cysteine proteases. All TS showed low toxicity (> 200μM) on cells of mammals. Although N4-methyl-4-nitrobenzaldehyde thiosemicarbazone, N4-methyl-4-nitroacetophenone thiosemicarbazone and its metal complex of manganese (Mn) were not active, the complex of Mn N4-methyl-4-nitrobenzophenone thiosemicarbazone revealed was moderately active on blood forms (IC50 19μM). The third class was DA, DNA binders with excellent activity against many pathogens. Although all have low toxicity cardiomyocyte, only three (DB1645, DB1582 and DB1651) were active against blood forms and intracellular (IC 50 from 0.15 to 13.3 mM), activity related to the curvature of the molecules (the molecules being the non-linear effective). OF all located in the nucleus and kDNA from parasites, with greater accumulation in the ulti
In vitro analysis of EIA DB766 on T.cruzi revealed: (i) excellent activity against bloodstream forms (60nm) and intracellular (25nm), while maintaining optimal action even when the parasite and incubated with 96% blood of mice, thus suggesting a potential prophylactic use, (ii) act on different strains (peridomestic / wild), including naturally resistant to benznidazole (BZ), with higher efficacy than BZ, (iii) location compartments rich DNA damage inducing ultrastructural the mitochondria . Based on the excellent selectivity index (> 533 and 714), tests were conducted during experimental acute infection by T. cruzi (Y and Colombian strains) in daily doses or alternate = 100mg/kg/day of DB766 alone or associated with BZ, administered ip or po for up to 20 days with the first dose at the onset of parasitaemia. DB766 (25 100mg/kg/day and 50mg/kg/day ip and po) reduced (> 90%) the parasite load (blood and cardiac tissue) and 90-100% protected against mortality, with similar efficacy to the BZ. EIA reversed inflammation and cardiac electrical changes and protected against liver injury induced by infection and muscle. Sub-optimal doses of BZ associated with DB289 (DA - pro-drug of DB75 - via po) or DB766 (ip) resulted in ≥ 99% reduction of parasitemia and 100% protective on mortality. BZ (po) + DB766 (ip) resulted in parasitological cure (2, 15 dogs) assessed by blood culture and PCR. EIA was the most active and selective against T. cruzi, encouraging the continuation of pre-clinical trials with representatives of this class in order to identification of new drugs for AD
| Identifer | oai:union.ndltd.org:IBICT/oai:www.arca.fiocruz.br:icict/4142 |
| Date | January 2009 |
| Creators | Batista, Denise da Gama Jaén |
| Contributors | Leon, Leonor Laura, Bahia, Maria Terezina, Barbosa, Helena Santos, Pereira, Mirian Claudia, Soeiro, Maria de Nazaré Correia |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Source | reponame:Repositório Institucional da FIOCRUZ, instname:Fundação Oswaldo Cruz, instacron:FIOCRUZ |
| Rights | info:eu-repo/semantics/openAccess |
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